Polypropylene binding peptides and methods of use

ABSTRACT

Combinatorially generated peptides are provided that have binding affinity for polypropylene (PP). The peptides may be used to deliver benefit agents to various PP surfaces.

CROSS-REFERENCE TO RELATED APPLICATION

This application claims priority under 35 U.S.C. §119 from U.S.Provisional Application Ser. No. 60/750,599, filed Dec. 15, 2005.

FIELD OF THE INVENTION

The invention relates to peptide based reagents having binding affinityfor polypropylene polymers.

BACKGROUND OF THE INVENTION

Polypropylene (PP) is a versatile thermoplastic resin available in awide range of formulations from general purpose homopolymer, randomcopolymer, and impact copolymer grades to highly specialized resins forengineering applications. Grades of polypropylene are available to meetthe needs of various fabrication processes such as extrusion, injectionmolding, thermoforming, blow molding, biaxially oriented film (BOPP),fiber spinning, slit tape, extrusion coating, and laminating.Polypropylene has good impact resistance and structural rigidity. It isunaffected by any solvent at room temperatures. It has excellentinsulating properties and is extremely lightweight. Its high fatiguestrength makes it a top choice under cyclic loading conditions. The widerange of physical properties and relative ease of processing makepolypropylene an extremely attractive material capable of competing withmore expensive resins in a number of demanding applications. For examplepolypropylene has very good resistance to fatigue, so that most plasticliving hinges, such as those on flip-top bottles, are made from thismaterial.

The ubiquitous use of PP in industry makes it a prime material candidatefor a variety of applications where the PP comprises some or all of asurface. One of the drawbacks to using PP as surface is that materialsthat bind to PP are specific and lack flexibility as binding agents. Sofor example where a new coating for PP is desired, a new search for a PPbinding molecule with the desired property must be conducted. Theresulting search is costly in both time and resources and not guaranteedto be successful. A system that is flexible and can be easily tailoredfor a variety of materials to be bound to PP is needed. The use ofpeptides as linkers or binders to PP offers some potential in thisregard.

Peptides having a binding affinity to polymer and plastic surfaces areknown. For example, Adey et al., (Gene 156:27-31 (1995)) describepeptides that bind to polystyrene and polyvinyl chloride surfaces.Additionally, peptides that bind to polyurethane (Murray et al., U.S.Patent Application Publication No. 2002/0098524), polyethyleneterephthalate (O'Brien et al., copending and commonly owned U.S. PatentApplication Publication No. 2005/0054752), and polystyrene,polyurethane, polycarbonate, and nylon (Grinstaff et al., U.S. PatentApplication Publication No. 2003/0185870) have been reported.

There remains a need therefore for a peptide based reagent that binds PPthat offers flexibility in bring a wide variety of materials to the PPsurface with minimum investment in redesign. Applicants have solved thestated problem by providing peptide reagents comprising PP bindingpeptides (PPBP). The PP binding peptides of the invention may bemodified with other functional or binding peptides allowing for thedelivery of benefit agents to the PP surface or for the use of thereagents to adhere PP containing surfaces.

SUMMARY OF THE INVENTION

The present invention provides PP binding peptides that may beincorporated into peptide based reagents useful for deliveringfunctional compounds to a PP surface. The PP binding peptides maycomprise active domains that have linker or other functionality ortarget binding domains that bind various benefit agents that aredelivered to the PP surface.

Accordingly, in one embodiment the invention provides a peptide reagenthaving a general structure selected from the group consisting of:

-   -   a) PP_(m)-(PPBP)_(n);    -   b) PP_(m)-(PPBP-BAp)n;    -   c) PP_(m)-(PPBP-AD)_(n);    -   d) PP_(m)-(PPBP-TBD)_(n); and    -   e) PP_(m)-(PPBP-L-BA)n; and    -   f) PP_(m)-[(PPBP)_(q)-L(x)-(PPBP)r]n-L-BA;    -   wherein:    -   i) PP is a PP moiety    -   ii) PPBP is a PP binding peptide having a PP binding domain;    -   iii) BA is at least one benefit agent;    -   iv) AD is at least one active domain incorporated into a PP        binding peptide;    -   v) TBD is at least one target binding domain incorporated into a        PP binding peptide;    -   vi) L is a linker molecule;    -   vii) m=the number of PP moieties available for binding;    -   viii) n=is less than or equal to m;    -   xi) p=1-20;    -   x) x=1-20; and    -   xi) r=1-50.        In an alternate embodiment the invention provides a peptide        reagent having the general structure:        (BA)_(n)-L_(m)-PP_(p)-[(X)_(a)-(Y)_(b)]_(q)-L_(r)-(BA)_(s)    -   Wherein:    -   i) BA is a benefit agent;    -   ii) PP is a PP moiety;    -   iii) L is a linker molecule;    -   iv) X is a PP peptide binding domain;    -   v) Y is an active domain; and    -   vi) wherein a, b, m, n, p, q, r, and s are non-negative integers        -   wherein, b, n, r, and s may be 0; and        -   a, p and q will at least be 1.

In another embodiment the invention provides a method for binding asubstrate comprising PP to a target comprising:

-   -   a) providing a peptide reagent of the invention: and    -   b) contacting the peptide reagent of (a) with a substrate        comprising a PP moiety under conditions whereby the peptide        reagent binds to the PP moiety.

Similarly the invention provides a method for delivering a benefit agentto a substrate comprising PP comprising:

-   -   a) providing the peptide reagent of the invention having a        benefit agent: and    -   b) contacting the peptide reagent of (a) with a substrate        comprising a PP moiety under conditions whereby the peptide        reagent binds to the PP moiety, whereby the benefit agent is        delivered to the substrate.

In an alternate embodiment the invention provides a method for adheringtwo surfaces comprising:

-   -   a) providing a first surface comprising PP comprising a first        peptide regent having the general formula;        (PPBP-AD1)        -   wherein:            -   i) PPBP is a PP binding peptide; and            -   ii) AD1 is a first active domain;    -   b) providing a second surface comprising a target molecule        comprising a second peptide reagent have the general formula;        (TBP-AD2)        -   wherein:            -   iii) TBP is a target binding peptide; and            -   iv) AD2 is a second active domain having affinity for                the first active domain;    -   c) juxtaposing the first and second surfaces wherein the first        and second peptide reagents adhere to each other through the        first and second active domains, whereby the surfaces are        adhered.

In a similar embodiment the invention provides a method for adhering twosurfaces comprising:

-   -   a) providing a first surface comprising a first target molecule        comprising a first peptide regent having the general formula;        (TBP1-AD)        -   wherein:            -   i) TBP1 is a first target binding peptide; and            -   ii) AD is an active domain having binding affinity for                PP;    -   b) providing a second surface comprising a second target        molecule comprising a second peptide reagent have the general        formula;        (TBP2-AD)        -   wherein:            -   iii) TBP2 is a second target binding peptide; and        -   iv) AD is an active domain having binding affinity for PP;    -   c) juxtaposing the first and second surfaces in the presence of        a PP moiety wherein the first and second peptide reagents adhere        to the PP moiety through the active domain, whereby the surfaces        are adhered.

Additionally the invention provides a PP binding peptide having an aminoacid sequence selected from the group consisting of SEQ ID NO's: 1-7.

BRIEF DESCRIPTION OF THE FIGURES AND SEQUENCE DESCRIPTIONS

FIG. 1 is a set of panels A-E which depict embodiments of the presentinvention as they are bound to a surface containing, in whole or inpart, PP particles.

FIG. 2 is a set of panels A-C which depict embodiments of the presentinvention as they are bound to a PP coating containing, in whole or inpart, PP particles, which is further bound to a surface.

FIG. 3 depicts an embodiment of the present invention as a diblock,optionally bound to a benefit agent at two different positions and/or atarget molecule. Also depicted is the optional inclusion of a linkermolecule and/or an active domain.

FIG. 4 depicts an embodiment of the present invention used to bond a PPcontaining surface with another surface which may contain PP or anotherknown target molecule.

FIG. 5 depicts an embodiment of the present invention used to bond tosurfaces together wherein neither necessarily contains PP.

FIG. 6 is a set of panels A-D which depict embodiments of the presentinvention used to coat a surface with PP.

SEQ ID NOs: 1-7 are PP binding sequences.

SEQ ID NOs:13-41 are antimicrobial peptides sequences.

SEQ ID NOs: 42-66 are pigment binding peptides sequences SEQ ID NOs:67-79 are print media binding peptide sequences:

SEQ ID NOs: 67 and 68 bind to cotton fabric, SEQ ID NOs: 67 and 69 bindto polyester/cotton fabric, SEQ ID NOs: 67, and 70-72 bind to HAMERMILL®paper, SEQ ID NOs: 74-78 bind to cellulose, and SEQ ID NO: 79 binds topoly(ethylene terephthalate).

SEQ ID NOs: 80-175 are body surface binding peptide sequences: SEQ IDNOs: 80-87 are skin-binding peptide sequences, SEQ ID NOs: 88-175 arehair binding peptide sequences and SEQ ID NOs: 88 and 89 bind nails aswell as hair.

SEQ ID NO:176 is the amino acid sequence of the Caspase 3 cleavage sitethat my be used as a peptide linker domain.

SEQ ID NOs: 177-179 are amino acid sequences of peptide linker domains.

A Sequence Listing is provided herewith on Compact Disk. The contents ofthe Compact Disk containing the Sequence Listing are hereby incorporatedby reference in compliance with 37 CFR 1.52(e). The Compact Disks aresubmitted in triplicate and are identical to one another. The disks arelabeled “Copy 1—Sequence Listing”, “Copy 2—Sequence Listing”, and CRF.The disks contain the following file: CL3312.ST25 having the followingsize: 39,000 bytes and which was created Nov. 20, 2006.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides variable coatings for PP substrates andsurfaces. More specifically, the present invention provides peptidesequences that bind PP with a high affinity. These peptides can be boundcovalently or otherwise to known substances to adapt PP for a variety ofuses. Additionally, the present invention provides methods to developand produce such peptides.

The following definitions and abbreviations are to be used for theinterpretation of the claims and the specification.

“BA” means benefit agent.

“PP” means polypropylene

“PPBP” is a PP binding peptide

“PBP” means pigment-binding peptide.

The term “peptide” refers to two or more amino acids joined to eachother by peptide bonds or modified peptide bonds.

The term “body surface” will mean any surface of the human body that mayserve as a substrate for the binding of a peptide carrying a benefitagent. Typical body surfaces include but are not limited to hair, skin,nails, teeth, gums, and corneal tissue.

The term “benefit agent” is a general term applying to a compound orsubstance that may be coupled with a complex of PP and PP bindingpeptide in order to provide a desirable characteristic of the benefitagent to the complex. In the most general sense a benefit agent may beany element, molecule or compound that is not PP or a PP-bindingpeptide. Benefit agents typically include colorants such as pigments anddyes as well as pharmaceuticals, markers, conditioners, and fragrances.

The term “hair” as used herein refers to human hair, eyebrows, andeyelashes.

The term “skin” as used herein refers to human skin, or pig skin,Vitro-Skin® and EpiDerm™ which are substitutes for human skin. Skin asused herein as a body surface will generally comprise a layer ofepithelial cells and may additionally comprise a layer of endothelialcells.

The term “nails” as used herein refers to human fingernails andtoenails.

The terms “coupling” and “coupled” as used herein refer to any chemicalassociation and includes both covalent and non-covalent interactions.

The term “pigment” refers to an insoluble, organic or inorganiccolorant.

The term “print medium” refers to any substrate suitable for printing.

The term “dispersant” as used herein refers to a substance thatstabilizes the formation of a colloidal solution of solid pigmentparticles in a liquid medium.

The term “PP binding peptide” refers to a peptide having specificaffinity for PP. The PP binding peptide will typically be short rangingfrom about 7 to about 50 amino acids in length and may be generatedrecombinantly, synthetically or may be selected by combinatorial means.PP binding peptides may comprise various subdomains including but notlimited to active domains, target domains and linker domains. Within anygiven PP binding peptide there resides a “PP binding domain” havingaffinity to PP. Any given PP binding peptide may contain only the PPbinding domain or may contain this domain in conjunction with active ortarget domains having different functionality.

The term “active domain” as used herein applies to a subsequence ofamino acids within a PP binding peptide. An active domain is a portionof the PP binding peptide that is not responsible for PP binding butprovides additional functionality or benefit. In one embodiment forexample an active domain may have antimicrobial functionality. In antherembodiment the active domain may have a linker function between twoother domains or between the peptide and a benefit agent. In anotherembodiment the active domain may serve to bind a specific target analyte(target domain).

The term “linking domain” or “linker domain” as used herein applies to aparticular of active domain that is used to either link two domainstogether, as a separator between two domains, or a domain and a terminalend. Linking domains may have a function beyond joining or separatingtwo features of a peptide.

The term “target binding domain” as used herein applies to a particulartype of peptide active domain that binds a target molecule, element,compound, or complex. The binding substrate for the target bindingdomain is referred to herein as the “target”. Typical targets willinclude but are not limited to biological analytes, (cells, cellmembrane fractions, viral proteins, proteins, antibodies, antibodyfragments, nucleic acids and the like), plant fibers, synthetic fibers,as well as organic and inorganic target complexes that will typically befound on surfaces or in print media. All target binding domains areactive domains. A “body surface binding domain” is a target domain thathas specific affinity for a body surface such as hair, skin, nails,teeth and the like. Similarly a “print media binding domain” willfunction to bind the elements of print media such as paper and other inkreceptive surfaces. Within the context of print media domains there maybe those domains that bind cellulose or cotton or other plant fibers.Additionally the target domains of the invention may be selected to bindspecific benefit agents such as colorants (pigments, dyes) andconditioners or any other organic or inorganic complex.

“PP moiety” means a discrete substance comprising polypropylene thatserves as a binding site for a PP binding peptide. PP moieties may makeup a PP film, or be comprised within various PP coatings on surfaces andsubstrates.

The term “linker” or “spacer” or ‘linker molecule” or “spacer molecule”will be used interchangeably and will mean a molecule or compound usedto bind a benefit agent to the PP-peptide complex. Any material that canbind said benefit agent to the complex can be used, including peptidebased molecules. A linker molecule is distinct from a linker domain inthat linker domains are inherently part of, or are proposed to be partof a peptide further comprising a PP-binding domain. A linker molecule,in whole or in part, may be identical to a linking domain, but a linkingmolecule does not contain a PP-binding domain.

As referred to herein a substance has “binding functionality” when itdemonstrates specific affinity for a substance or target.

As referred to herein a substance has “catalytic functionality” when itdemonstrates the ability to catalyze a chemical reaction

As referred to herein a substance has “antimicrobial functionality whenit demonstrates the ability to kill microbial cell populations.

As used herein the term “surface” when used in conjunction with a PPmoiety means the point of contact for the PP moiety. Surfaces of theinvention will typically be coated with PP or may themselves comprise PPmoieties. Surfaces may take the form of solid support, a bead, amicrosphere, a sheet, or a fiber. In some instances the surface of theinvention may be layered or juxtaposed on a “secondary surface”. A“secondary surface” will typically be coated or layers with the PPsurfaces of the invention.

The term “diblock structure” as used herein refers to a composition thatconsists of two different units or blocks, each serving a specificfunction. The peptide-based diblock polymers of the present inventionconsist of a PP-binding peptide block coupled to a substrate, or aPP-binding peptide block coupled to a benefit agent. The diblock polymermay contain multiple copies of the peptide block.

The term “triblock structure” as used herein refers to a pigmentdispersant that consists of three different units or blocks, eachserving a specific function. The peptide-based triblock structure of thepresent invention consists of a substrate-block, PP-binding peptideblock, and a benefit agent block. The triblock structure may containmultiple copies of any of the peptide blocks.

The term “stringency” as it is applied to the selection of PP bindingpeptides, hair-binding, skin-binding, and nail-binding peptides of thepresent invention, refers to the concentration of the eluting agent(usually detergent) used to elute peptides from the substrate to whichthey are bound or for which they have affinity. Higher concentrations ofthe eluting agent provide more stringent conditions.

The term “amino acid” refers to the basic chemical structural unit of aprotein or polypeptide. The following abbreviations are used herein toidentify specific amino acids:

Three-Letter One-Letter Amino Acid Abbreviation Abbreviation Alanine AlaA Arginine Arg R Asparagine Asn N Aspartic acid Asp D Cysteine Cys CGlutamine Gln Q Glutamic acid Glu E Glycine Gly G Histidine His HIsoleucine Ile I Leucine Leu L Lysine Lys K Methionine Met MPhenylalanine Phe F Proline Pro P Serine Ser S Threonine Thr TTryptophan Trp W Tyrosine Tyr Y Valine Val V

“Gene” refers to a nucleic acid fragment that expresses a specificprotein, including regulatory sequences preceding (5′ non-codingsequences) and following (3′ non-coding sequences) the coding sequence.“Native gene” refers to a gene as found in nature with its ownregulatory sequences “Chimeric gene” refers to any gene that is not anative gene, comprising regulatory and coding sequences that are notfound together in nature. Accordingly, a chimeric gene may compriseregulatory sequences and coding sequences that are derived fromdifferent sources, or regulatory sequences and coding sequences derivedfrom the same source, but arranged in a manner different than that foundin nature. A “foreign” gene refers to a gene not normally found in thehost organism, but that is introduced into the host organism by genetransfer. Foreign genes can comprise native genes inserted into anon-native organism, or chimeric genes.

“Synthetic genes” can be assembled from oligonucleotide building blocksthat are chemically synthesized using procedures known to those skilledin the art. These building blocks are ligated and annealed to form genesegments which are then enzymatically assembled to construct the entiregene. “Chemically synthesized”, as related to a sequence of DNA, meansthat the component nucleotides were assembled in vitro. Manual chemicalsynthesis of DNA may be accomplished using well-established procedures,or automated chemical synthesis can be performed using one of a numberof commercially available machines. Accordingly, the genes can betailored for optimal gene expression based on optimization of nucleotidesequence to reflect the codon bias of the host cell. The skilled artisanappreciates the likelihood of successful gene expression if codon usageis biased towards those codons favored by the host. Determination ofpreferred codons can be based on a survey of genes derived from the hostcell where sequence information is available.

“Coding sequence” refers to a DNA sequence that codes for a specificamino acid sequence. “Suitable regulatory sequences” refer to nucleotidesequences located upstream (5′ non-coding sequences), within, ordownstream (3′ non-coding sequences) of a coding sequence, and whichinfluence the transcription, RNA processing or stability, or translationof the associated coding sequence. Regulatory sequences may includepromoters, translation leader sequences, introns, polyadenylationrecognition sequences, RNA processing site, effector binding site andstem-loop structure.

“Promoter” refers to a DNA sequence capable of controlling theexpression of a coding sequence or functional RNA. In general, a codingsequence is located 3′ to a promoter sequence. Promoters may be derivedin their entirety from a native gene, or be composed of differentelements derived from different promoters found in nature, or evencomprise synthetic DNA segments. It is understood by those skilled inthe art that different promoters may direct the expression of a gene indifferent tissues or cell types, or at different stages of development,or in response to different environmental or physiological conditions.Promoters which cause a gene to be expressed in most cell types at mosttimes are commonly referred to as “constitutive promoters”. It isfurther recognized that since in most cases the exact boundaries ofregulatory sequences have not been completely defined, DNA fragments ofdifferent lengths may have identical promoter activity.

The term “expression”, as used herein, refers to the transcription andstable accumulation of sense (mRNA) or antisense RNA derived from thenucleic acid fragment of the invention. Expression may also refer totranslation of mRNA into a polypeptide.

The term “transformation” refers to the transfer of a nucleic acidfragment into the genome of a host organism, resulting in geneticallystable inheritance. Host organisms containing the transformed nucleicacid fragments are referred to as “transgenic” or “recombinant” or“transformed” organisms.

The term “host cell” refers to cell which has been transformed ortransfected, or is capable of transformation or transfection by anexogenous polynucleotide sequence.

The terms “plasmid”, “vector” and “cassette” refer to an extrachromosomal element often carrying genes which are not part of thecentral metabolism of the cell, and usually in the form of circulardouble-stranded DNA molecules. Such elements may be autonomouslyreplicating sequences, genome integrating sequences, phage or nucleotidesequences, linear or circular, of a single- or double-stranded DNA orRNA, derived from any source, in which a number of nucleotide sequenceshave been joined or recombined into a unique construction which iscapable of introducing a promoter fragment and DNA sequence for aselected gene product along with appropriate 3′ untranslated sequenceinto a cell. “Transformation cassette” refers to a specific vectorcontaining a foreign gene and having elements in addition to the foreigngene that facilitate transformation of a particular host cell.“Expression cassette” refers to a specific vector containing a foreigngene and having elements in addition to the foreign gene that allow forenhanced expression of that gene in a foreign host.

The term “phage” or “bacteriophage” refers to a virus that infectsbacteria. Altered forms may be used for the purpose of the presentinvention. The preferred bacteriophage is derived from the “wild” phage,called M13. The M13 system can grow inside a bacterium, so that it doesnot destroy the cell it infects but causes it to make new phagescontinuously. It is a single-stranded DNA phage.

The term “phage display” refers to the display of functional foreignpeptides or small proteins on the surface of bacteriophage or phagemidparticles. Genetically engineered phage may be used to present peptidesas segments of their native surface proteins. Peptide libraries may beproduced by populations of phage with different gene sequences.

Standard recombinant DNA and molecular cloning techniques used hereinare well known in the art and are described by Sambrook, J., Fritsch, E.F. and Maniatis, T., Molecular Cloning: A Laboratory Manual, SecondEdition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.(1989) (hereinafter “Maniatis”); and by Silhavy, T. J., Bennan, M. L.and Enquist, L. W., Experiments with Gene Fusions, Cold Spring HarborLaboratory Cold Press Spring Harbor, N.Y. (1984); and by Ausubel, F. M.et al., Current Protocols in Molecular Biology, published by GreenePublishing Assoc. and Wiley-Interscience (1987).

The present invention relates to peptides and peptide reagents that havespecific binding affinity to PP in various conformations includingcomplexes of the PP binding peptides linked to benefit agents, andoptionally where the PP binding peptides comprise active peptide domainsor target binding domains having binding or other functionality forother substances or surfaces. The PP peptide regent of the invention maytake a variety of forms including those represented by the followingstructures:

-   -   a) PP_(m)-(PPBP)_(n);    -   b) PP_(m)-(PPBP-BAp)n;    -   c) PP_(m)-(PPBP-AD)_(n);    -   d) PP_(m)-(PPBP-TBD)_(n); and    -   e) PP_(m)-(PPBP-L-BA)n; and    -   f) PP_(m)-[(PPBP)_(q)-L(x)-(PPBP)r]n-L-BA;

wherein:

-   -   i) PP is a PP moiety    -   ii) PPBP is a PP binding peptide having a PP binding domain;    -   iii) BA is at least one benefit agent;    -   iv) AD is at least one active domain incorporated into a PP        binding peptide;    -   v) TBD is at least one target binding domain incorporated into a        PP binding peptide;    -   vi) L is a linker molecule;    -   vii) m=the number of PP moieties available for binding;    -   viii) n=is less than or equal to m;    -   xi) p=1-20;    -   x) x=1-20; and    -   xi) r=1-50.

Alternatively the PP peptides reagents of the invention may beconfigured according formula:(BA)_(n)-L_(m)-PP_(p)-[(X)_(a)-(Y)_(b)]_(q)-L_(r)-(BA)_(s)

-   -   Wherein:    -   i) BA is a benefit agent;    -   ii) PP is a PP moiety;    -   iii) L is a linker molecule;    -   iv) X is a PP peptide binding domain;    -   v) Y is an active domain; and    -   vi) wherein a, b, m, n, p, q, r, and s are non-negative integers        -   wherein, b, n, r, and s may be 0; and        -   a, p and q will at least be 1.            Polypropylene Moieties

A polypropylene (PP) moiety, as defined herein, is the binding site of apolypropylene-binding peptide on a surface. Typically it is expectedthat surface area of the PP moiety that is presented to the PP bindingpeptide will be on the order of about 200 Angstroms to about 5000angstroms.

The PP moiety may be incorporated into a surface in various ways. Forexample, the surface may be the surface of a PP substrate.Alternatively, the PP moiety may be imbedded into the surface of anothermaterial, such as a polymer, or may be a film or coating on the surfaceof another material, such as a metal, polymer, glass, cloth, and thelike. The PP moiety may comprise a PP polymer or a copolymer of propenewith other α-olefins, such as ethylene and butene. The copolymer may bea random or block copolymer.

Polypropylene is prepared commercially by the polymerization of propene,typically with a heterogeneous Ziegler-Natta catalyst or a homogeneouscatalyst, such as a metallocene, using suspension, bulk, solution or gasphase polymerization processes (Ullmann's Encyclopedia of IndustrialChemistry, 6^(th) edition, 2003, Wiley-VCH Verlag GmbH and Co.,Weinheim, Germany, Vol. 28, pp. 432-445). The PP polymer may beprocessed into various shapes or forms, such as beads, microspheres,sheets, rods, tubes, films, plates, rings, fiber, yarns, andmicrofilament, using injection molding, blow molding, extrusion,extrusion coating and roll extrusion techniques, which are well known inthe art. Polypropylene and polypropylene copolymers in various shapesare available commercially from companies such as Equistar Chemicals, LP(Houston, Tex.), Huntsman Corp. (Salt Lake City, Utah), Blueridge Films,Inc. (McKenney, Va.), and Trident Plastics, Inc. (Ivyland, Pa.).

In one embodiment, the PP polymer or copolymer is a film bonded toanother surface, such as metal, polymer, glass, cloth, and the like,using methods known in the art, such as heat sealing.

In another embodiment, the PP polymer or copolymer is a coating onanother surface, such as metal, polymer, glass, cloth, and the like,prepared using methods known in the art, such as extrusion coating.

In another embodiment, the PP polymer or copolymer is imbedded into thesurface of another material, such as another polymer. This may be doneby adding particles, beads, or fragments of PP material into the otherpolymer as it cures or crystallizes.

Identification of PP-Binding Peptides

Peptides having affinity for PP, referred to herein as PP-bindingpeptides (PPBP), are peptide sequences that bind strongly to a PPmoiety. The PP-binding peptides of the invention are from about 7 aminoacids to about 50 amino acids, more preferably, from about 7 amino acidsto about 25 amino acids, most preferably from about 7 to about 20 aminoacids in length. Suitable PP-binding peptides may be selected usingmethods that are well known in the art.

The PP-binding peptides may be generated randomly and then selectedagainst a PP substrate based upon their binding affinity for PP, asdescribed by O'Brien et al. (copending and commonly owned U.S. PatentApplication Publication No. 2005/0054752), Adey et al., (Gene 156:27-31,(1995)), Murray et al. (U.S. Patent Application Publication No.2002/0098524) and Grinstaff et al. (U.S. Patent Application PublicationNo. 2003/0185870), all of which are incorporated herein by reference.The generation of random libraries of peptides is well known and may beaccomplished by a variety of techniques including, bacterial display(Kemp, D. J.; Proc. Natl. Acad. Sci. USA 78(7):4520-4524 (1981), andHelfman et al., Proc. Natl. Acad. Sci. USA 80(1):31-35, (1983)), yeastdisplay (Chien et al., Proc Natl Acad Sci USA 88(21):9578-82 (1991)),combinatorial solid phase peptide synthesis (U.S. Pat. No. 5,449,754,U.S. Pat. No. 5,480,971, U.S. Pat. No. 5,585,275, U.S. Pat. No.5,639,603), and phage display technology (U.S. Pat. No. 5,223,409, U.S.Pat. No. 5,403,484, U.S. Pat. No. 5,571,698, U.S. Pat. No. 5,837,500).Techniques to generate such biological peptide libraries are well knownin the art. Exemplary methods are described in Dani, M., J. of Receptor& Signal Transduction Res., 21 (4):447-468 (2001), Sidhu et al., Methodsin Enzymology 328:333-363 (2000), and Phage Display of Peptides andProteins, A Laboratory Manual, Brian K. Kay, Jill Winter, and JohnMcCafferty, eds.; Academic Press, NY, 1996. Additionally, phage displaylibraries are available commercially from companies such as New EnglandBioLabs (Beverly, Mass.).

A preferred method to randomly generate peptides is by phage display.Phage display is an in vitro selection technique in which a peptide orprotein is genetically fused to a coat protein of a bacteriophage,resulting in display of fused peptide on the exterior of the phagevirion, while the DNA encoding the fusion resides within the virion.This physical linkage between the displayed peptide and the DNA encodingit allows screening of vast numbers of variants of peptides, each linkedto a corresponding DNA sequence, by a simple in vitro selectionprocedure called “biopanning”. In its simplest form, biopanning iscarried out by incubating the pool of phage-displayed variants with atarget of interest that has been immobilized on a plate or bead, washingaway unbound phage, and eluting specifically bound phage by disruptingthe binding interactions between the phage and the target. The elutedphage is then amplified in vivo and the process is repeated, resultingin a stepwise enrichment of the phage pool in favor of the tightestbinding sequences. After 3 or more rounds of selection/amplification,individual clones are characterized by DNA sequencing.

Specifically, the PP-binding peptides may be selected using thefollowing method. A suitable library of phage-peptides is generatedusing the methods described above or the library is purchased from acommercial supplier. After the library of phage-peptides has beengenerated, they are then contacted with an appropriate amount of thepolymer substrate. The library of phage-peptides is dissolved in asuitable solution for contacting the substrate. The test substrate maybe suspended in the solution or may be immobilized on a plate or bead. Apreferred solution is a buffered aqueous saline solution containing asurfactant. A suitable solution is Tris-buffered saline (TBS) with 0.5%Tween® 20. The solution may additionally be agitated by any means inorder to increase the mass transfer rate of the peptides to the polymersubstrate, thereby shortening the time required to attain maximumbinding.

Upon contact, a number of the randomly generated phage-peptides willbind to the polymer substrate to form a phage-peptide-polymer complex.Unbound phage-peptide may be removed by washing. After all unboundmaterial is removed, phage-peptides having varying degrees of bindingaffinities for the polymer substrate may be fractionated by selectedwashings in buffers having varying stringencies. Increasing thestringency of the buffer used increases the required strength of thebond between the phage-peptide and polymer substrate in thephage-peptide-substrate complex.

A number of substances may be used to vary the stringency of the buffersolution in peptide selection including, but not limited to, acidic pH(1.5-3.0); basic pH (10-12.5); high salt concentrations such as MgCl₂(3-5 M) and LiCl (5-10 M); water; ethylene glycol (25-50%); dioxane(5-20%); thiocyanate (1-5 M); guanidine (2-5 M); urea (2-8 M); andvarious concentrations of different surfactants such as SDS (sodiumdodecyl sulfate), DOC (sodium deoxycholate), Nonidet P-40, Triton X-100,Tween® 20, wherein Tween® 20 is preferred. These substances may beprepared in buffer solutions including, but not limited to, Tris-HCl,Tris-buffered saline, Tris-borate, Tris-acetic acid, triethylamine,phosphate buffer, and glycine-HCl, wherein Tris-buffered saline solutionis preferred.

It will be appreciated that phage-peptides having increasing bindingaffinities for the PP substrate may be eluted by repeating the selectionprocess using buffers with increasing stringencies. The elutedphage-peptides can be identified and sequenced by any means known in theart.

In one embodiment, the following method for generating the PP-bindingpeptides of the present invention may be used. A library ofcombinatorially generated phage-peptides is contacted with PP to formphage peptide-substrate complexes. The phage-peptide-substrate complexis separated from uncomplexed peptides and unbound substrate, and thebound phage-peptides from the phage-peptide-substrate complexes areeluted from the complex, preferably by acid treatment. Then, the elutedphage-peptides are identified and sequenced. To identify peptidesequences that bind to PP but not to other substrates, a subtractivepanning step may be added. Specifically, the library of combinatoriallygenerated phage-peptides is first contacted with the non-target toremove phage-peptides that bind to it. Then, the non-bindingphage-peptides are contacted with PP and the above process is followed.Alternatively, the library of combinatorially generated phage-peptidesmay be contacted with the non-target and PP simultaneously. Then, thephage-peptide-substrate complexes are separated from thephage-peptide-non-target complexes and the method described above isfollowed for the desired phage-substrate complexes.

Alternatively, a modified phage display screening method for isolatingpeptides with a higher affinity for polymer substrates may be used. Inthe modified method, the phage-peptide-substrate complexes are formed asdescribed above. Then, these complexes are treated with an elutionbuffer. Any of the elution buffers described above may be used.Preferably, the elution buffer is an acidic solution. Then, theremaining, elution-resistant phage-peptide-substrate complexes are usedto directly infect/transfect a bacterial host cell, such as E. coliER2738. The infected host cells are grown in an appropriate growthmedium, such as LB (Luria-Bertani) medium, and this culture is spreadonto agar, containing a suitable growth medium, such as LB medium withIPTG (isopropyl β-D-thiogalactopyranoside) and S-Gal™. After growth, theplaques are picked for DNA isolation and sequencing to identify thepeptide sequences with a high binding affinity for the substrate ofinterest. Alternatively, PCR may be used to identify theelution-resistant phage-peptides from the modified phage displayscreening method, described above, by directly carrying out PCR on thephage-peptide-substrate complexes using the appropriate primers, asdescribed by Janssen et al. in U.S. Patent Application Publication No.2003/0152976, which is incorporated herein by reference.

Production of PP-Binding Peptides

The PP-binding peptides of the present invention may be prepared usingstandard peptide synthesis methods, which are well known in the art (seefor example Stewart et al., Solid Phase Peptide Synthesis, PierceChemical Co., Rockford, Ill., 1984; Bodanszky, Principles of PeptideSynthesis, Springer-Verlag, New York, 1984; and Pennington et al.,Peptide Synthesis Protocols, Humana Press, Totowa, N.J., 1994).Additionally, many companies offer custom peptide synthesis services.

Alternatively, the PP-binding peptides of the present invention may beprepared using recombinant DNA and molecular cloning techniques. Genesencoding the PP-binding peptides may be produced in heterologous hostcells, particularly in the cells of microbial hosts, as described byHuang et al. (U.S. Patent Application Publication No. 2005/0050656) andO'Brien et al., supra.

Preferred heterologous host cells for expression of the binding peptidesof the present invention are microbial hosts that can be found broadlywithin the fungal or bacterial families and which grow over a wide rangeof temperature, pH values, and solvent tolerances. Becausetranscription, translation, and the protein biosynthetic apparatus arethe same irrespective of the cellular feedstock, functional genes areexpressed irrespective of carbon feedstock used to generate cellularbiomass. Examples of host strains include, but are not limited to,fungal or yeast species such as Aspergillus, Trichoderma, Saccharomyces,Pichia, Candida, Hansenula, or bacterial species such as Salmonella,Bacillus, Acinetobacter, Rhodococcus, Streptomyces, Escherichia,Pseudomonas, Methylomonas, Methylobacter, Alcaligenes, Synechocystis,Anabaena, Thiobacillus, Methanobacterium and Klebsiella.

A variety of expression systems can be used to produce the peptides ofthe present invention. Such vectors include, but are not limited to,chromosomal, episomal and virus-derived vectors, e.g., vectors derivedfrom bacterial plasmids, from bacteriophage, from transposons, frominsertion elements, from yeast episoms, from viruses such asbaculoviruses, retroviruses and vectors derived from combinationsthereof such as those derived from plasmid and bacteriophage geneticelements, such as cosmids and phagemids. The expression systemconstructs may contain regulatory regions that regulate as well asengender expression. In general, any system or vector suitable tomaintain, propagate or express polynucleotide or polypeptide in a hostcell may be used for expression in this regard. Microbial expressionsystems and expression vectors contain regulatory sequences that directhigh level expression of foreign proteins relative to the growth of thehost cell. Regulatory sequences are well known to those skilled in theart and examples include, but are not limited to, those which cause theexpression of a gene to be turned on or off in response to a chemical orphysical stimulus, including the presence of regulatory elements in thevector, for example, enhancer sequences. Any of these can be used toconstruct chimeric genes for production of the any of the bindingpeptides of the present invention. These chimeric genes can then beintroduced into appropriate microorganisms via transformation to providehigh level expression of the peptides.

Vectors or cassettes useful for the transformation of suitable hostcells are well known in the art. Typically the vector or cassettecontains sequences directing transcription and translation of therelevant gene, one or more selectable markers, and sequences allowingautonomous replication or chromosomal integration. Suitable vectorscomprise a region 5′ of the gene, which harbors transcriptionalinitiation controls and a region 3′ of the DNA fragment which controlstranscriptional termination. It is most preferred when both controlregions are derived from genes homologous to the transformed host cell,although it is to be understood that such control regions need not bederived from the genes native to the specific species chosen as aproduction host. Selectable marker genes provide a phenotypic trait forselection of the transformed host cells such as tetracycline orampicillin resistance in E. coli.

Initiation control regions or promoters which are useful to driveexpression of the chimeric gene in the desired host cell are numerousand familiar to those skilled in the art. Virtually any promoter capableof driving the gene is suitable for producing the binding peptides ofthe present invention including, but not limited to: CYC1, HIS3, GAL1,GAL10, ADH1, PGK, PHO5, GAPDH, ADC1, TRP1, URA3, LEU2, ENO, TPI (usefulfor expression in Saccharomyces); AOX1 (useful for expression inPichia); and lac, ara, tet, trp, IP_(L), IP_(R), T7, tac, and trc(useful for expression in Escherichia coli) as well as the amy, apr, nprpromoters and various phage promoters useful for expression in Bacillus.

Termination control regions may also be derived from various genesnative to the preferred hosts. Optionally, a termination site may beunnecessary, however, it is most preferred if included.

The vector containing the appropriate DNA sequence as described supra,as well as an appropriate promoter or control sequence, may be employedto transform an appropriate host to permit the host to express thepeptide of the present invention. Cell-free translation systems can alsobe employed to produce such peptides using RNAs derived from the DNAconstructs of the present invention. Optionally, it may be desired toproduce the instant gene product as a secretion product of thetransformed host. Secretion of desired proteins into the growth mediahas the advantages of simplified and less costly purificationprocedures. It is well known in the art that secretion signal sequencesare often useful in facilitating the active transport of expressibleproteins across cell membranes. The creation of a transformed hostcapable of secretion may be accomplished by the incorporation of a DNAsequence that codes for a secretion signal which is functional in theproduction host. Methods for choosing appropriate signal sequences arewell known in the art (see for example EP 546049 and WO 9324631). Thesecretion signal DNA or facilitator may be located between theexpression-controlling DNA and the instant gene or gene fragment, and inthe same reading frame with the latter.

Active Domains

As noted above active domains are peptide portions of the PP bindingpeptide that convey various additional functionality to the peptide. Anysequence of amino acids may be used as an active domain, including, butnot limited to those functioning as a linker, those having bindingfunctionality, having catalytic functionality and those havingantimicrobial functionality.

An antimicrobial active domain may be particularly desirable if the PPmoiety part of the diblock was for instance part of a kitchen countertopsurface. Such antimicrobial sequences are well known in the art. Anypeptide based antimicrobial sequence could be used as an active domainin the above embodiment. As non-limiting examples table 1 providespossible antimicrobial active domain sequences.

TABLE 1 Antimicrobial active domain sequences. SEQ Species of ID originNO. Sequence Artificial 13 PKGLKKLLKGLKKLLKL Artificial 14KGLKKLLKGLKKLLKL Artificial 15 KGLKKLLKLLKKLLKL Artificial 16LKKLLKLLKKLLKL Artificial 17 LKKLLKLLKKLL Artificial 18VAKKLAKLAKKLAKLAL Artificial 19 FAKLLAKALKKLL Artificial 20KGLKKGLKLLKKLLKL Artificial 21 KGLKKLLKLGKKLLKL Artificial 22KGLKKLGKLLKKLLKL Artificial 23 KGLKKLLKLLKKGLKL Artificial 24KGLKKLLKLLKKLGKL Artificial 25 FALALKALKKLKKALKKAL Artificial 26FAKKLAKLAKKLAKLAL Artificial 27 FAKLLAKLAKKLL Artificial 28FAKKLAKLALKLAKL Artificial 29 FAKKLAKKLL Artificial 30 FAKLLAKLAKKVLArtificial 31 KYKKALKKLAKLL Artificial 32 FALLKALLKKAL Artificial 33KRLFKKLKFSLRKY Artificial 34 KRLFKKLLFSLRKY Artificial 35 LLLFLLKKRKKRKYH. cecropia 36 KWKLFKKIEKVGQNIRDGIIKAGPAVAWGQATQIAK Xenopus 37GIGKFLHSAKKFGKAFVGEIMNS Xenopus 38 GIGKFLKKAKKFGKAFVKILKK Bos Taurus 39RLCRIVVIRVCR Bos Sp. 40 ILPWKWPWWPWRR H. sapiens 41DSHAKRHHGYKRKFHEKHHSHRGY

Two sub-types of active domains, target binding domains and linkingdomains, have been given specific names in the discussion of thispresent invention. A target binding domain is an active domain thatspecifically binds to a known target. Target binding sequences are knownin the art and can be developed using known techniques as well astechniques described herein. Non-limiting examples of targets to whichtarget binding domains will bind include, pigments, dyes, chemicalfunctional groups, print media, body surfaces (hair, skin, nails, teethetc.) and biological analytes (cells, receptors, proteins, nucleicacids, viral particles, prions etc.) (see FIGS. 4, 5 and 6 panel D).

A linking domain is an active domain that is specifically used toseparate two domains or a domain from a terminal end. Any sequence ofamino acids that does not contain a PP-binding site can be used as alinking domain. A linking domain can have activity beyond justseparating two features of a peptide. A linking domain may provide aspecific structure to the separating portion of the peptide. Conversely,a linking domain may also be selected to provide flexibility to theseparating portion of the peptide. Additionally the linking domain maybe created to specifically change the rheology of the medium the peptideis immersed in. Also the linking domain may be constructed so that itcan be cleaved by, or act as the binding site for, a cleaving moleculeor enzyme, for the purpose of releasing a portion of the peptide and/orthe PP from the complex.

Preferred peptide linker domains are composed of the amino acidsproline, lysine, glycine, alanine, and serine, and mixtures thereof. Inaddition, the peptide linker may contain a specific enzyme cleavagesite, such as the protease Caspase 3 site, given by SEQ ID NO:176, whichallows for the enzymatic removal of a portion of the peptide and/or thePP from the complex. The peptide linker may be from 1 to about 50 aminoacids, preferably from 1 to about 20 amino acids in length. Examples ofpeptide linkers include, but are not limited to, SEQ ID NOs:176 to 179.These peptide linkers may be linked to the binding peptide sequence byany method know in the art. For example, the entire bindingpeptide-peptide linker-diblock may be prepared using the standardpeptide synthesis methods described supra. In addition, the bindingpeptide and peptide linker blocks may be combined using carbodiimidecoupling agents (see for example, Hermanson, Bioconjugate Techniques,Academic Press, New York (1996)), diacid chlorides, diisocyanates andother difunctional coupling reagents that are reactive to terminal amineand/or carboxylic acid groups on the peptides. Alternatively, the entirebinding peptide-peptide linker-diblock may be prepared using therecombinant DNA and molecular cloning techniques described supra. Thelinker may also be a combination of a peptide linker and an organiclinker molecule, which may be prepared using the methods describedabove. Examples of specific linker peptides are given in table 2 below.

TABLE 2 Linker peptides SEQ Species of ID origin NO. Sequence Artificial176 LESGDEVD Artificial 177 TSTSKASTTT TSSKTTTTSS KTTTTTSKTS TTSSSSTArtificial 178 GQGGYGGLGS QGAGRGGLGG QG Artificial 179 GPGGYGPGQQ

Target domains of the invention are another type of active domaincomprised within the PP binding peptide. Target domains will havebinding affinity for various substance such as benefit agents (pigments,dyes, print media, biological analtyes, body surfaces (hair, skin,nails, teeth etc.) and the like).

Pigment binding domains are target domains that bind various pigmentsand colorants. Such pigments have application in the personal care aswell as the printing industries. Similarly print media binding domainsare target binding domains having specific affinity for various types ofprint media. Typically the print media will comprise cotton or cellulosetargets or may be coated with a polymer such as nylon or PP giving riseto cotton, cellulose or polymer binding domains as part of the PPbinding peptide.

Target domains may be uni-functional having binding affinity for asingle target species or multifunctional, having affinity for a varietyof targets. For example it may be desirable to combine a pigment bindingdomain or a print medium binding domain or both into the peptide part ofthe PP-peptide complex of the present invention. Such an embodiment thatincludes a print-medium binding domain may be particularly desirable ifthe complex already contains a benefit agent that is a colorant or dye.Pigment-binding peptides and print medium-binding peptides have beenidentified (See tables 3, 4, and 5, and O'Brien et al., supra, herebyincorporated by reference. The pigment-binding peptides typicallycomprise at least about 40 mole % of the amino acids: glycine, alanine,valine, leucine, isoleucine, methionine, proline, phenylalanine, andtryptophan Specifically, binding peptides were isolated that have a highaffinity for the pigments carbon black, given as SEQ ID NOs:42-45,CROMOPHTHAL® Yellow, given as SEQ ID NOs: 46-53, SUNFAST® Magenta, givenas SEQ ID NOs: 55-57, and SUNFAST® Blue, given as SEQ ID NOs: 54, 58-66.The cellulose-binding peptides of the invention comprise at least about14 mole % of the amino acids: serine, threonine and tyrosine. Bindingpeptides having a high binding affinity for cellulose (a major componentof cotton) include SEQ ID NOs: 73-78. The polyester-binding peptides ofthe invention comprise at least about 20 mole % of the amino acids:phenylalanine, tryptophan, and tyrosine. Binding peptides having a highaffinity for polyester (poly(ethylene terephthalate)) include SEQ ID NO:79. Additionally, binding peptides were isolated that have a bindingaffinity for the following print media: cotton, given as SEQ ID NOs: 67and 68, polyester/cotton, given as SEQ ID NOs: 67 and 69, and printingpaper, given as SEQ ID NOs: 67, and 70-72.

TABLE 3 Pigment-Binding Peptides SEQ Designated Peptide ID Pigment NameSequence NO: Carbon Black CB-71 MPPPLMQ 42 CB-72 FHENWPS 43 CB-121RTAPTTPLLLSL 44 CB-122 WHLSWSPVPLPT 45 Cromophtal ® CY-71 PHARLVG 46Yellow CY-72 NIPYHHP 47 CY-73 TTMPAIP 48 CY-74 HNLPPRS 49 CY-121AHKTQMGVRQPA 50 CY-122* ADNVQMGVSHTP 51 CY-123* AHNAQMGVSHPP 52 CY-124*ADYVGMGVSHRP 53 CY-125 SVSVGMKPSPRP 54 Sunfast ® Magenta SM-71 YPNTALV55 SM-72 VATRIVS 56 SM-121 HSLKNSMLTVMA 57 Sunfast ® Blue SB-71 NYPTQAP58 SB-72 KCCYSVG 59 SB-121 RHDLNTWLPPVK 60 SB-122 EISLPAKLPSAS 61 SB-123SVSVGMKPSPRP 54 SB-124** SDYVGMRPSPRH 62 SB-125** SDYVGMRLSPSQ 63SB-126** SVSVGIQPSPRP 64 SB-127** YVSVGIKPSPRP 65 SB-128** YVCEGIHPCPRP66 *These sequences are analogs of CY-121. **These sequences are eitheranalogs of SB-123 or are similar to the analogs of SB-123.

TABLE 4 Print Medium-Binding Peptides SEQ Designated Peptide IDPrint Medium Name Sequence NO: Cotton fabric COT-71* SILPYPY 67 COT-72STASYTR 68 Polyester/cotton fabric P/C-71 LPVRPWT 69 P/C-72* SILPYPY 67Hammermill ® paper HCP-71 GNTPSRA 70 HCP-72 HAIYPRH 71 HCP-73 YQDSAKT 72HCP-74* SILPYPY 67 *These sequences are identical.

TABLE 5 Cellulose and Poly(ethylene terephthalate)- Binding PeptidesPrint Medium Designated Peptide SEQ ID Ingredient Name Sequence NO:Cellulose CEL-71 VPRVTSI 73 CEL-72 MANHNLS 74 CEL-73 FHENWPS 75 CEL-121THKTSTQRLLAA 76 CEL-122 KCCYVNVGSVFS 77 CEL-123 AHMQFRTSLTPH 78Poly(ethylene PET-121 GTSDHMIMPFFN 79 terephthalate)

Target domains that have binding affinity for body surfaces areparticularly useful for the production of personal care compositionscomprising colorants, and conditioners with specific binding affinityfor the body surface. For example, it may be desirable to attachPP-peptide complex of the present invention in either a tri-block formor a di-block form to a body surface such as hair or skin. One method toachieve such a result is to incorporate a target binding domain into thepeptide part of the present invention that binds hair, skin or anotherbody surface. Both hair and skin binding domains can be produced by themethods described here, in the co-pending, commonly owned U.S. Ser. No.10/935,642 (U.S. Patent Application Publication No. 2005/0050656) herebyincorporated by reference and in co-pending, commonly owned U.S. Ser.No. 11/074,473 (U.S. Patent Application Publication No. 2005/0226839)also hereby incorporated by reference. Examples of hair and skin bindingdomains are shown in Table 6.

TABLE 6 Body Surface Binding Peptide Domains SEQ Body ID Surface NOSequence Skin 80 FTQSLPR Skin 81 TPFHSPENAPGS Skin 82 KQATFPPNPTAY Skin83 HGHMVSTSQLSI Skin 84 LSPSRMK Skin 85 LPIPRMK Skin 86 HQRPYLT Skin 87FPPLLRL Nail 88 ALPRIANTWSPS Nail 89 YPSFSPTYRPAF Hair 90 YPSFSPTYRPAFHair 91 ALPRIANTWSPS Hair 92 LESTPKMK Hair 93 SVSVGMKPSPRP Hair 94LDVESYKGTSMP Hair 95 RVPNKTVTVDGA Hair 96 DRHKSKYSSTKS Hair 97KNFPQQKEFPLS Hair 98 QRNSPPAMSRRD Hair 99 TRKPNMPHGQYL Hair 100KPPHLAKLPFTT Hair 101 NKRPPTSHRIHA Hair 102 NLPRYQPPCKPL Hair 103RPPWKKPIPPSE Hair 104 RQRPKDHFFSRP Hair 105 SVPNK(T or P)VTVDGX Hair 106TTKWRHRAPVSP Hair 107 WLGKNRIKPRAS Hair 108 SNFKTPLPLTQS Hair 109KELQTRNVVQRE Hair 110 GMPAMHWIHPFA Hair 111 TPTANQFTQSVP Hair 112AAGLSQKHERNR Hair 113 ETVHQTPLSDRP Hair 114 LPALHIQRHPRM Hair 115QPSHSQSHNLRS Hair 116 RGSQKSKPPRPP Hair 117 THTQKTPLLYYH Hair 118TKGSSQAILKST Hair 119 DLHTVYH Hair 120 HIKPPTR Hair 121 HPVWPAI Hair 122MPLYYLQ Hair 123 HLTVPWRGGGSAVPFYSHSQITLPNH Hair 124GPHDTSSGGVRPNLHHTSKKEKRENRKVPFYSHSVTSRG NV Hair 125 KHPTYRQ Hair 126HPMSAPR Hair 127 MPKYYLQ Hair 128 MHAHSIA Hair 129 TAATTSP Hair 130LGIPQNL Hair 131 AKPISQHLQRGS Hair 132 APPTPAAASATT Hair 133DPTEGARRTIMT Hair 134 EQISGSLVAAPW Hair 135 LDTSFPPVPFHA Hair 136LPRIANTWSPS Hair 137 RTNAADHPAAVT Hair 138 SLNWVTIPGPKI Hair 139TDMQAPTKSYSN Hair 140 TIMTKSPSLSCG Hair 141 TPALDGLRQPLR Hair 142TYPASRLPLLAP Hair 143 AKTHKHPAPSYS Hair 144 TDPTPFSISPER Hair 145CAAGCCTCAGCGACCGAATA Hair 146 WHDKPQNSSKST Hair 147 NEVPARNAPWLV Hair148 NSPGYQADSVAIG Hair 149 TQDSAQKSPSPL Hair 150 TPPELLHGDPRS Hair 151TPPTNVLMLATK Hair 152 NTSQLST Hair 153 NTPKENW Hair 154 NTPASNR Hair 155PRGMLST Hair 156 PPTYLST Hair 157 TIPTHRQHDYRS Hair 158 TPPTHRL Hair 159LPTMSTP Hair 160 LGTNSTP Hair 161 TPLTGSTNLLSS Hair 162 TPLTKET Hair 163QQSHNPP Hair 164 TQPHNPP Hair 165 STNLLRTSTVHP Hair 166 HTQPSYSSTNLFHair 167 SLLSSHA Hair 168 QQSSISLSSHAV Hair 169 NASPSSL Hair 170 HSPSSLRHair 171 K(H, R or N)SHHTH Hair 172 E(H, R, or N)SHHTH Hair 173 LESTSLLHair 174 TPLTKET Hair 175 KQSHNPP

If the present invention is desired to be used in connection with a haircare composition an effective amount of the complex for use in a haircare composition is herein defined as a proportion of from about 0.01%to about 10%, preferably about 0.01% to about 5% by weight relative tothe total weight of the composition. Components of a cosmeticallyacceptable medium for hair care compositions are described by Philippeet al. in U.S. Pat. No. 6,280,747, and by Omura et al. in U.S. Pat. No.6,139,851 and Cannell et al. in U.S. Pat. No. 6,013,250, all of whichare incorporated herein by reference. For example, these hair carecompositions can be aqueous, alcoholic or aqueous-alcoholic solutions,the alcohol preferably being ethanol or isopropanol, in a proportion offrom about 1 to about 75% by weight relative to the total weight, forthe aqueous-alcoholic solutions. Additionally, the hare carecompositions may contain one or more conventional cosmetic ordermatological additives or adjuvants including but not limited to,antioxidants, preserving agents, fillers, surfactants, UVA and/or UVBsunscreens, fragrances, thickeners, wetting agents and anionic, nonionicor amphoteric polymers, and dyes or pigments.

In a number of embodiments the present invention could be used in a skincare composition. Skin care compositions are herein defined ascompositions comprising an effective amount of a skin conditioner or amixture of different skin conditioners in a cosmetically acceptablemedium. The uses of these compositions include, but are not limited to,skin care, skin cleansing, make-up, and anti-wrinkle products. If thepresent invention is desired to be used in connection with a skin carecomposition an effective amount of the complex for skin carecompositions is herein defined as a proportion of from about 0.001% toabout 10%, preferably about 0.01% to about 5% by weight relative to thetotal weight of the composition. This proportion may vary as a functionof the type of skin care composition. Suitable compositions for acosmetically acceptable medium are described by Philippe et al. supra.For example, the cosmetically acceptable medium may be an anhydrouscomposition containing a fatty substance in a proportion generally offrom about 10 to about 90% by weight relative to the total weight of thecomposition, where the fatty phase containing at least one liquid, solidor semi-solid fatty substance. The fatty substance includes, but is notlimited to, oils, waxes, gums, and so-called pasty fatty substances.Alternatively, the compositions may be in the form of a stabledispersion such as a water-in-oil or oil-in-water emulsion.Additionally, the compositions may contain one or more conventionalcosmetic or dermatological additives or adjuvants, including but notlimited to, antioxidants, preserving agents, fillers, surfactants, UVAand/or UVB sunscreens, fragrances, thickeners, wetting agents andanionic, nonionic or amphoteric polymers, and dyes or pigments.

Benefit Agents

Benefit agents are any material or substance that may be complexed withthe PP binding peptide in an manner so as to deliver a benefit at thepoint where the PP binding peptide is attached. In the most generalsense the benefit agent will be the third component of the tri-block ofPP and PP binding peptide. Any complex, compound or element may be usedwith the present invention as a benefit agent. If a user of theinvention desires to have the features of a benefit agent combined withPP then a triblock may be constructed to include the benefit agent inthe formation with PP and a PP-binding peptide. A benefit agent may beselected for the purpose of adding the physical, chemical and/orbiological properties of said agent to the PP-peptide complex of thepresent invention. The result of this construct will be said benefitagent closely associated with PP and the activity of said benefit agentwill be included within the triblock.

The triblock embodiment of this present invention is composed of atleast one member of each block element but may also have multiple copiesof identical or different members of one, two or all three blockelements. Benefit agents can be used singularly or in a plurality. Insome embodiments a plurality of peptide blocks or a plurality of PPblocks or a plurality of both blocks may be added to a single benefitagent block or a number of benefit agent blocks. For some small benefitagents, for non-limited example, those composed of an element, as manyas 10,000 benefit agents could be added to a single PP-complex. For somelarge benefit agents, for non-limiting example, a dye embedded in aplastic bead as many as 10,000 PP complexes might be attached to asingle benefit agent.

Benefit agents may be inorganic or organic in nature, this includesbeing polymer or peptide based. They may not be by definition eithercomposed of PP or part of a PP-binding peptide, since such compositionsare defined as categorically other parts of the triblock formation. Somepreferred embodiments include benefit agents that are pigments,pharmaceuticals, markers, conditioners, colorants, and fragrances.

Pharmaceuticals.

A pharmaceutical generally means a substance dosed to an organism orthing to treat or prevent a disease or condition. A pharmaceuticalbenefit agent includes, in a non-limiting sense, the topical, internalor intracellular administration of a compound to an organism as atreatment or a prophylactic. A non-limiting example of this embodimentof the present invention would be the attachment of an anti-acnemedication to formulation of the present invention designed to be a skinconditioner. A pharmaceutical benefit agent also includes a treatment tosurface or item to prevent an infectious germ from being transmittedafter contacting said surface or item. The addition of an antimicrobialcompound to a construction of the present invention to be used oncountertops would be a non-limiting example of this embodiment. Suitablepharmaceuticals are well known in the art. An extensive list is given byKabonov et al. in U.S. Pat. No. 6,696,089, which is incorporated hereinby reference (in particular, columns 16 to 18). Examples include, butare not limited to, antibacterial agents, antiviral agents, antifungalagents, anti-cancer agents, vaccines, radiolabels, anti-inflammatories,anti-glaucomic agents, local anesthetics, anti-neoplastic agents,antibodies, hormones, and the like.

Markers.

Markers as used and defined herein refer to a class of benefit agentsthat provide aid in detecting the presence of the PP-peptide complex towhich they are, or were, attached. The marker benefit agent might be adye, fluorescent label, radioactive element or some other signal.Radioactive P³² is a non-limiting example of this type of marker benefitagent. Also the marker benefit agent might also be a substance thatreacts with a dye, fluorescent label or other signal. Biotin used inconnection with a labeled-streptavidin compound is a non-limited exampleof this type of marker benefit agent. Additionally a marker benefitagent might also provide, or help to provide aid to detect, the presenceor lack of presence of another specific chemical, compound, element orcomplex. By way of non-limiting example, the marker benefit agent mightbe a compound that is metabolized by a specific enzyme to produce ametabolite that reacts with a fluorescently labeled phosphine. TheStaudinger ligation is a non-limiting example of this type of markerbenefit agent.

Conditioners.

Conditioner benefits agents as referred to in discussion of the presentinvention generally mean benefit agents that provide an improvement tothe appearance, texture or quality of the substance they are designed tocondition. Conditioner benefit agents may be used with the presentinvention to condition any substance including but not limited to hair,skin, lips, leather, and upholstery. In the preferred embodiment thepresent invention is used in combination with a benefit agent thatprovides a conditioning effect to hair and skin. In the most preferredembodiment said hair and skin are human hair and human skin.

Hair conditioning agents as herein defined are agents which improve theappearance, texture, and sheen of hair as well as increasing hair bodyor suppleness. In the peptide-based hair conditioners of the presentinvention, any known hair conditioning agent may be used. Hairconditioning agents are well known in the art, see for example Green etal. (WO 0107009), incorporated herein by reference, and are availablecommercially from various sources. Suitable examples of hairconditioning agents include, but are not limited to, cationic polymers,such as cationized guar gum, diallyly quaternary ammoniumsalt/acrylamide copolymers, quaternized polyvinylpyrrolidone andderivatives thereof, and various polyquaternium-compounds; cationicsurfactants, such as stearalkonium chloride, centrimonium chloride, andSapamin hydrochloride; fatty alcohols, such as behenyl alcohol; fattyamines, such as stearyl amine; waxes; esters; nonionic polymers, such aspolyvinylpyrrolidone, polyvinyl alcohol, and polyethylene glycol;silicones; siloxanes, such as decamethylcyclopentasiloxane; polymeremulsions, such as amodimethicone; and volumizing agents, such asnanoparticles (e.g., silica nanoparticles and polymer nanoparticles).The preferred hair conditioning agents of the present invention containamine or hydroxyl functional groups to facilitate coupling to thehair-binding peptides. Examples of preferred conditioning agents areoctylamine (CAS No. 111-864), stearyl amine (CAS No. 124-30-1), behenylalcohol (CAS No. 661-19-8, Cognis Corp., Cincinnati, Ohio), vinyl groupterminated siloxanes, vinyl group terminated silicone (CAS No.68083-19-2), vinyl group terminated methyl vinyl siloxanes, vinyl groupterminated methyl vinyl silicone (CAS No. 68951-99-5), hydroxylterminated siloxanes, hydroxyl terminated silicone (CAS No. 80801-30-5),amino-modified silicone derivatives, [(aminoethyl)amino]propyl hydroxyldimethyl siloxanes, [(aminoethyl)amino]propyl hydroxyl dimethylsilicones, and alpha-tridecyl-omega-hydroxy-poly(oxy-1,2-ethanediyl)(CAS No. 24938-91-8).

Skin conditioning agents as herein defined include, but are not limitedto astringents, which tighten skin; exfoliants, which remove dead skincells; emollients, which help maintain a smooth, soft, pliableappearance; humectants, which increase the water content of the toplayer of skin; occlusives, which retard evaporation of water from theskin's surface; and miscellaneous compounds that enhance the appearanceof dry or damaged skin or reduce flaking and restore suppleness. In thepeptide-based skin conditioners of the present invention, any known skinconditioning agent may be used. Skin conditioning agents are well knownin the art, see for example Green et al. supra, and are availablecommercially from various sources. Suitable examples of skinconditioning agents include, but are not limited to, alpha-hydroxyacids, beta-hydroxy acids, polyols, hyaluronic acid, D,L-panthenol,polysalicylates, vitamin A palmitate, vitamin E acetate, glycerin,sorbitol, silicones, silicone derivatives, lanolin, natural oils andtriglyceride esters. The preferred skin conditioning agents of thepresent invention are polysalicylates, propylene glycol (CAS No.57-55-6, Dow Chemical, Midland, Mich.), glycerin (CAS No. 56-81-5,Proctor & Gamble Co., Cincinnati, Ohio), glycolic acid (CAS No. 79-14-1,DuPont Co., Wilmington, Del.), lactic acid (CAS No. 50-21-5, Alfa Aesar,Ward Hill, Mass.), malic acid (CAS No. 617-48-1, Alfa Aesar), citricacid (CAS No. 77-92-9, Alfa Aesar), tartaric acid (CAS NO. 133-37-9,Alfa Aesar), glucaric acid (CAS No. 87-73-0), galactaric acid (CAS No.526-99-8), 3-hydroxyvaleric acid (CAS No. 10237-77-1), salicylic acid(CAS No. 69-72-7, Alfa Aesar), and 1,3 propanediol (CAS No. 504-63-2,DuPont Co., Wilmington, Del.). Polysalicylates may be prepared by themethod described by White et al. in U.S. Pat. No. 4,855,483,incorporated herein by reference. Glucaric acid may be synthesized usingthe method described by Merbouh et al. (Carbohydr. Res. 336:75-78(2001). The 3-hydroxyvaleric acid may be prepared as described byBramucci in WO 02012530.

Colorants.

The term colorant generally refers to a coloring agent. Colorants may bechemically organic or inorganic and may include pigments or dyes. Thepeptide-based colorants of the present invention may be prepared bycovalently attaching a specific PP-binding peptide to a coloring agent,either directly or via a linker, using any of the coupling methods knownin the art (see for example, U.S. Patent Application Publication No.2005/0226839).

Pigments are a particularly suitable benefit agent. Pigments generallymeans an insoluble colorant. A wide variety of organic and inorganicpigments alone or in combination may be used in the present invention.Examples of organic pigments include, but are not limited to Cyan,Yellow, Red, Blue, Orange, Magenta, Black, Green, Violet, Light Cyan,and Light Magenta. Preferred organic pigments are carbon black, such asCarbon Black FW18, and colored pigments such as CROMOPHTHAL® Yellow131AK (Ciba Specialty Chemicals), SUNFAST® Magenta 122 (Sun Chemical)and SUNFAST® Blue 15:3 (Sun Chemical). Examples of inorganic pigmentsinclude, but are not limited to finely divided metals, such as copper,iron, aluminum, and alloys thereof; and metal oxides, such as silica,alumina, and titania. Additional examples of suitable pigments are givenby Ma et al. in U.S. Pat. No. 5,085,698, incorporated herein byreference.

The preferred coloring agents for use in the skin based applications ofthe present invention include but are not limited to the following dyes:eosin derivatives such as D&C Red No. 21 and halogenated fluoresceinderivatives such as D&C Red No. 27, D&C Red Orange No. 5 in combinationwith D&C Red No. 21 and D&C Orange No. 10, and the pigments: titaniumdioxide, zinc oxide, D&C Red No. 36 and D&C Orange No. 17, the calciumlakes of D&C Red Nos. 7, 11, 31 and 34, the barium lake of D&C Red No.12, the strontium lake D&C Red No. 13, the aluminum lakes of FD&C YellowNo. 5, of FD&C Yellow No. 6, of D&C Red No. 27, of D&C Red No. 21, ofFD&C Blue No. 1, iron oxides, manganese violet, chromium oxide,ultramarine blue, and carbon black.

The preferred coloring agents for use with the present invention in thenail based applications include but are not limited to D&C Red Nos. 8,10, 30 and 36, the barium lakes of D&C Red Nos. 6, 9 and 12, the calciumlakes of D&C Red Nos. 7, 11, 31 and 34, the strontium lake of D&C RedNo. 30 and D&C Orange No. 17 and D&C Blue No. 6. Suitable hair coloringagents for use with the present invention include, but are not limitedto dyes, such as 4-hydroxypropylamino-3-nitrophenol,4-amino-3-nitrophenol, 2-amino-6-chloro-4-nitrophenol,2-nitro-paraphenylenediamine, N,N-hydroxyethyl-2-nitro-phenylenediamine,4-nitro-indole, Henna, HC Blue 1, HC Blue 2, HC Yellow 4, HC Red 3, HCRed 5, Disperse Violet 4, Disperse Black 9, HC Blue 7, HC Blue 12, HCYellow 2, HC Yellow 6, HC Yellow 8, HC Yellow 12, HC Brown 2, D&C Yellow1, D&C Yellow 3, D&C Blue 1, Disperse Blue 3, Disperse violet 1, eosinderivatives such as D&C Red No. 21 and halogenated fluoresceinderivatives such as D&C Red No. 27, D&C Red Orange No. 5 in combinationwith D&C Red No. 21 and D&C Orange No. 10; and pigments, such as D&C RedNo. 36 and D&C Orange No. 17, the calcium lakes of D&C Red Nos. 7, 11,31 and 34, the barium lake of D&C Red No. 12, the strontium lake of D&CRed No. 13, the aluminum lakes of FD&C Yellow No. 5, of FD&C Yellow No.6, of D&C Red No. 27, of D&C Red No. 21, and of FD&C Blue No. 1, ironoxides, manganese violet, chromium oxide, titanium dioxide, iron oxides,zinc oxide, barium oxide, ultramarine blue, bismuth citrate, and carbonblack particles.

The preferred hair coloring agents of the present invention are D&CYellow 1 and 3, HC Yellow 6 and 8, D&C Blue 1, HC Blue 1, HC Brown 2, HCRed 5, 2-nitro-paraphenylenediamine,N,N-hydroxyethyl-2-nitro-phenylenediamine, 4-nitro-indole, iron oxides,and carbon black.

Fragrances.

A fragrance is a complex, compound or element that releases, a substancewhich may be perceived by the sense of olfaction or chemical detectionin any organism, but preferably, in humans. The object sensed ordetected may be a part of or the whole of the fragrance benefit agent.In the preferred embodiment the odor is perceived as desirable tohumans. However, some uses may combine the present invention with afragrance benefit agent that is repellent to a class of organisms,including a class that contains or is humans. Any known fragrance orodor may be use as a benefit agent. It may be desirable to attach afragrance benefit agent to the PP-peptide complex by a bond structure orlinking molecule that allows the benefit agent to be released, in partor in whole, so that it may be perceived by a sensing organ or chemicaldetector.

Numerous fragrances, both natural and synthetic, are well known in theart. For example, Secondini (Handbook of Perfumes and Flavors, ChemicalPublishing Co., Inc., New York, 1990), incorporated herein by reference,describes many of the natural and synthetic fragrances used incosmetics. Suitable natural fragrances include, but are not limited, tojasmines, narcissus, rose, violet, lavender, mint, spice, vanilla,anise, amber, orange, pine, lemon, wintergreen, rosemary, basil, andspruce. Suitable synthetic fragrances include, but are no limited to,acetaldehyde, C7 to C16 alcohols, benzyl acetate, butyric acid, citricacid, isobutyl phenyl acetate, linalyl butyrate, malic acid, menthol,phenyl ethyl cinnamate, phenyl propyl formate, tannic acid, terpineol,vanillin, amyl salicylate, benzaldehyde, diphenyl ketone, indole, andthe like.

Linker Molecules

Linker molecules may optionally be used with some embodiments of thepresent invention for the purpose of attaching the benefit agent to thePP-peptide complex (see FIG. 3, reference number 225). Any molecule,compound or complex that will attach the benefit agent to the complexcan be used a linking molecule provided the linking molecule is does notcontain PP or a PP-binding domain. The benefit agent may be attached tothe complex to either the PP moiety or the peptide portion or in thecase of a plurality of benefit agent possibly to both. The linkingmolecules may be designed to bond the benefit agent stably or in thealternative they may be designed to break and release the benefit agentfrom the complex in a given circumstance. Such circumstances could be,for non-limiting example, a range of pH, a range of temperatures, arange of pressure, while immersed in a certain media, the presence of aparticular element, molecule or compound at a certain range ofconcentration, after a given passage of time, or at a certain averagerate for a population of linker molecules.

Specifically the linker may be any of a variety of molecules, such asalkyl chains, phenyl compounds, ethylene glycol, amides, esters and thelike. Preferred linkers are hydrophilic and have a chain length from 1to about 100 atoms, more preferably, from 2 to about 30 atoms. Examplesof preferred linkers include, but are not limited to, ethanol amine,ethylene glycol, polyethylene with a chain length of 6 carbon atoms,polyethylene glycol with 3 to 6 repeating units, phenoxyethanol,propanolamide, butylene glycol, butyleneglycolamide, propyl phenyl, andethyl, propyl, hexyl, steryl, cetyl, and palmitoyl alkyl chains. Thelinker may be covalently attached to the peptide and the benefit agentusing any of the coupling chemistries described above. In order tofacilitate incorporation of the linker, a bifunctional cross-linkingagent that contains a linker and reactive groups at both ends forcoupling to the peptide and the benefit agent may be used. Suitablebifunctional cross-linking agents are well known in the art and include,but are not limited to diamines, such a as 1,6-diaminohexane;dialdehydes, such as glutaraldehyde; bis N-hydroxysuccinimide esters,such as ethylene glycol-bis(succinic acid N-hydroxysuccinimide ester),disuccinimidyl glutarate, disuccinimidyl suberate, and ethyleneglycol-bis(succinimidylsuccinate); diisocyantes, such ashexamethylenediisocyanate; bis oxiranes, such as 1,4 butanediyldiglycidyl ether; dicarboxylic acids, such as succinyldisalicylate; andthe like. Heterobifunctional cross-linking agents, which contain adifferent reactive group at each end, may also be used.

Applications of PP Binding Peptides

It will be appreciated by the skilled person that PP binding peptidescomprising active and target domains having specific functionality maybe used in a multiplicity of formats including as delivery means fordelivering benefits agents, in assays for diagnostic applications aswell as in materials applications for coating PP surfaces. The followingdescription of the figures presents a limited number of examples of themethod of the invention, but is by no means inclusive of all possibleapplications and formats.

Referring to FIG. 1 panel A, there is shown a surface 1 comprising, inwhole or in part, a PP moiety 3. At least some of the PP molecules 3 areexposed in various orientations on the exterior of the surface. The PPbinding peptide 5 comprises at least one, but not limited to one, PPbinding domain 7. The PP binding domain 7 further comprises at leastone, but not limited to one, PP binding site 15. PP binding peptides 5will bind specifically to PP molecules 3, this binding will occur at thePP binding site 15 of the PP domain 7 within the PP binding peptide 5.The PP-binding peptide 5 coupled to the PP moiety 3 forms a diblockstructure. The formation of this diblock structure on a PP containingsurface 1, such as CORIAN® is useful for coating such surfaces with aprotein layer to serve as a sacrificial layer or to mask properties ofthe surface 1.

FIG. 1 panel B depicts another embodiment of the invention. In thisembodiment, a PP binding peptide 5 also binds to a surface 1 comprising,in whole or in part, PP molecules 3. A benefit agent 19 is coupled tothe PP binding peptide 5 covalently, ionically or otherwise as describedelsewhere herein. Although bound to the PP binding peptide 5, thebenefit agent 19 generally retains the biological, chemical and physicalproperties that it exhibited before being coupled to the PP bindingpeptide 5. The complex of the PP particle 3, the PP-binding peptide 5,and the benefit agent 19 forms a triblock. The proximity of the benefitagent 19 to the surface 1 after binding allows the benefit agent 19 tobe active at that location, and provides the chemical property of thebenefit agent 19 on the PP containing surface 1. Non-limiting examplesof the benefit agents 19 are colorants such as dyes and pigments,conditioners, fragrances, pharmaceuticals and the like.

FIG. 1 panel C depicts still another embodiment of the invention. The PPbinding peptide 5 binds to a surface 1 comprising PP 3 as above. PanelC, as in panels A and B, shows the PP binding peptide 5 comprising atleast one, but not limited to one, PP binding domain 7 within itsstructure. The PP binding domain 7 comprises at least one, but notlimited to one, PP binding site 15. The PP binding peptide 5 of panel Cfurther comprises at least one, but not limited to one, active domain 9different from the PP binding domain 7, yet within the same PP bindingpeptide 5. By having an active domain 9 within the peptide 5 and thepeptide 5 being bound to a PP-containing surface 1 this embodiment ofthe invention allows the property of the active domain 9 to betransmitted to the surface 1. One non-limiting example of an activedomain as exemplified here is a domain having antimicrobial properties.

FIG. 1 panel D depicts still another embodiment of the invention. The PPbinding peptide 5 binds to a surface 1 comprising PP 3 as describedabove. In this embodiment, the PP binding peptide 5 comprises a specifictarget binding domain 11 targeting other molecules other than PP. Inthis embodiment, the PP-binding peptide 1 acts as an intermediary tobring the target molecules close to the surface 1. This may be used toprovide the chemical, biologic or physical function of target molecule17 on the surface 1. However, this embodiment may also be employed toisolate the target molecule 17 from the surrounding media. Another usemay be to sample the surrounding media for the presence of the targetmolecule 17. Non-limiting examples of the other target molecule 17include benefit agents such as colorants (dyes and pigments) andconditioners as well as biological analytes, (cells, membrane fractions,viral particles, proteins, nucleic acids and the like), body surfaces,(hair, skin, nails, teeth and the like) as well as other organic andinorganic target complexes.

FIG. 1 panel E depicts still another embodiment of the invention. The PPbinding peptide 5 binds to a surface 1 comprising PP 3 as above. Panel Edepicts a PP binding peptide 5 that contains a linker domain 13 thatserves to connect the PP binding peptide 5 to a benefit agent 19. Thelinker domain 13 is a domain that selected to physically separate thebenefit agent 19 from the PP binding domain(s) 7. Alternatively,although not depicted in the FIG. 1, a single linker domain or manylinker domains may be provided to separate various domains within thePP-binding peptide. For instance, it may be advantageous to separate thePP binding domain form an active domain, or to separate two or moreactive domains, a linker could be utilized to achieve this separation.The linker domain 13 may simply provide a steric benefit. Although insome uses of this embodiment the linker provides a specific structure ororientation between the PP binding peptide 5 and the benefit agent 19 orto limit the conformation of the benefit agent-PP binding peptide-PPtriblock. In other uses of this embodiment the linker 13 provides aflexible region so that the benefit agent-PP binding peptide-PP triblockcan form a particular conformation or a variety of differentconformations. Still, in other uses of this embodiment the chemical andphysical nature of the linker 13 may be used to change the rheology ofthe environment surrounding the surface 1 to which the peptide 5 isbound. Non-limiting examples of linker domains 13 that would alter therheology of the surrounding surface include, hydrophobic, hydrophilic,or charged molecules. Additionally a linker domain 13 may be employed torelease a benefit agent 19 from the PP-binding peptide 5 under variouscircumstances. Such circumstances may include for example, a certainrange of pH, or a certain range of temperatures, or a certain range ofpressures. Such circumstances may also include response to shock,response the presence of a particular molecule, especially a peptidecleaving molecule, or the passage of time.

Referring to FIG. 2 panel A, a surface 101 is shown that could becomprised of any surface material. Non-limiting examples of suchsurfaces are metal, paper, glass and cloth. A coating 121 comprised ofin whole or in part, PP molecules or moieties 103 has been applied tothe surface 101 as shown. In this embodiment of the invention, aPP-binding peptide 105 is targeted to the coating. As in the abovedescriptions the PP-binding peptide 105 comprises, in whole or in part,at least one PP-binding domain 107, which itself comprises, in whole orin part, at least one PP-binding site 115. As described elsewhere hereinthe PP-binding site 115 binds specifically to PP molecules 103. In thisembodiment the PP-binding site 115 binds to exposed portions of PP 103in a PP coating 121 on attached to the surface 101. In this embodimentthe PP-binding peptide 105 is useful to provide an additional coating toPP coating 103 already applied to the surface 101. Non-limiting examplesof the uses for this embodiment include a sacrificial layer to protectthe PP coating or in the case of multiple PP domains 107 and/or bindingsites 115 to act as an adhesive between the PP coat 121 and other PPmoieties 103 or surfaces 101.

Similar to Panel A, FIG. 2 Panel B depicts a PP-binding peptide 105coupled to a PP coating 121 on a surface 101. The PP-binding peptide 105depicted in panel B further comprises a benefit agent 119. In thisembodiment of the invention, at least one, but not limited to one,benefit agent 119 is coupled to the PP-binding peptide 105 by acovalent, ionic or other interactive means. The PP bindingpeptide-benefit agent complex is in turn coupled to a PP moiety 103within the PP coating 121 on the surface 101. As described elsewhereherein the benefit agent 119 has some activity or functionally thatpersists while it is bound to the PP-binding peptide 105 and the complexis bound to the PP coating 121. The active benefit agent 119 beingbrought to or near the coating 121 by the PP-binding peptide 105 conveysits activity to the coating, modifies the coating or enhances thecoating. In a similar embodiment, a plurality of different types ofPP-binding peptides 105 may be used in combination so that the differentbenefit agents 119 corresponding to the different PP-binding peptides105 may interact, or act in concert to produce a desirable result.

Similar to Panel A, FIG. 2 Panel C depicts a PP-binding peptide 105bound to a PP coating 121 on a surface 101. The PP-binding peptide 105comprises all of the features of the PP-binding peptide 105 depicted inPanel A, with the added feature that the peptide includes an activedomain 109 separate from the PP-binding domain. This active domain 109,remains active as part of the PP-binding peptide 105, and furthercontinues to be active when the PP-binding peptide 105 binds to PP 103within the PP coating 121. The active domain 109, by virtue of beingpart of the diblock containing the PP-binding peptide 105 and the PPcoating 121, is active in the proximity of the coating 121. This allowsthe coating 121 to exhibit the activity of the active domain 109contained in the PP-binding peptide 105 bound to the PP molecules 103contained within the coating 121. Additionally, as in panel B, a benefitagent 119 may also be chemically attached to the functional domaincontaining PP-binding peptide 105 (not shown). As stated above, anadditional embodiment would be the use of a plurality of different typesof PP-binding peptides 105 to interact or act in concert to produce adesirable result.

Referring to FIG. 3, another embodiment of the invention is shown inwhich the PP substrate is not bound to a larger surface. In thisembodiment the PP exists as a PP moiety 203 which may be suspended insolution, such as cell growth media, air, water, oil, biological fluids,gels and the like. A PP-binding peptide 205 is depicted bound to the PPmoiety 203. The PP-binding peptide 205 contains within its peptidestructure at least one, but not limited to one PP binding domain 207.The PP binding domain 207 contains within its structure a PP bindingsite 215. PP binding domains 207 and PP binding sites 215 bind PPmoieties 203 specifically as described elsewhere herein. Binding of thePP-binding peptide 205 to the PP 203 occurs at the PP binding site 215with the PP binding domain 207. The binding of PP moieties 203 to thePP-binding peptide 205 forms a diblock structure.

Other embodiments of the invention add additional elements to thedi-block formation of PP moiety 203 and PP-binding peptide 205. One suchembodiment, involves binding a benefit agent 219 to the PP-bindingpeptide 205. The addition of a benefit agent 219 to the diblockstructure forms a triblock structure. The benefit agent 219 may becoupled to the PP-binding peptide 205 by any known means, as describedabove. The function of benefit agents 219 is discussed in greater detailelsewhere herein.

Alternatively the benefit agent 223 may be attached to the PP moiety203. In this format, the benefit agent 223 is attached to the PP moietyor bead 203 typically by chemical means or bonds 225. The bond may bepart of the benefit agent 223 or may be an independent structure that isbound to the PP 203 for the purpose of binding the benefits agent 223.In the alternative, the bond structure 225 may be bound to the benefitagent 223 for the purpose of binding it to the PP 203. The bindingstructure 225 may be a permanent bond, but in some forms of theembodiment may be easily broken under certain conditions. In other formsof the embodiment, the bond 225 may allow the benefit agent 223 to beleached from the PP moiety or bead 203 under certain conditions. Instill other forms of the embodiment, the bond 225 may allow the benefitagent 223 to be released over time at regular or specific timeintervals. Alternatively, the bond 225 itself may be in whole or in partbe composed of PP. In this way, the PP moiety or bead 203 may be inwhole or in part the binding structure 225. The benefit agent 223 may bepartially or fully embedded with the PP moiety or bead 203.

In another embodiment described by FIG. 3, the PP-binding peptide 205 isbound the PP 203 as described above and the complex may optionally bebound to a benefit agent 219 and/or 223 by any method describedelsewhere herein. The additional feature of this embodiment is at leastone or a plurality of additional active peptide domains 209 within thePP-binding peptide 205. Any known peptide active domain 209 can be usedin this embodiment. Alternatively, the active domain 209 may be a linkerdomain or may function as a target domain and bind a target 217.

Additional embodiments of the invention are illustrated in FIGS. 4 and5. Referring to FIGS. 4 and 5, a plurality of PP-binding peptides 305may be employed to bring substances together. FIG. 4 depicts aPP-binding peptide 305 used to bring a PP containing surface 301together with another surface 333. The PP containing surface 301 iscomprised in whole or in part of PP moieties 303 At least some PPmoieties 303 are exposed in part or in full on the at least one side ofthe surface 301. Likewise, the non-PP surface or complementary surface333, is comprised in whole or in part of a known moiety 335 for whichthere exists a peptide binding-domain 329 or for which a peptide bindingdomain 329 can be designed using the methods described elsewhere herein.The amino acid structure of the PP-binding peptides 305 comprises inwhole or in part of at least one PP binding domain 307, but possiblymore that one. The PP binding domain 307 itself comprises of at leastone or a plurality of PP binding sites 315. PP binding sites 315 areable to bind to the exposed PP 303 of the PP containing surface 301 andin such way to adhere the PP-binding peptides 305 to surface 301. Inaddition to comprising of one or more PP binding domains 307, thePP-binding peptide 305 of this embodiment also comprises of at leastone, but possibly more, target binding domains 311. The target bindingdomain 311 specifically binds another target domain 337 in a handshakefashion allowing the complex to serve as an adhesive binding the PP andnon-PP containing surfaces together. The target binding domain 311 insome uses may be capable of binding to itself. In that case, the targetbinding domain 311 and the target domain 337 could be identical.

The complementary surface 333 is composed a known surface-exposed moietyor a complementary moiety 335, for which there is a known peptidebinding domain 329, a complementary peptide binding domain 329. Acomplementary moiety binding peptide 327 is composed of at least one,but possibly more than one, complementary moiety binding domain 329,which itself is composed of at least one but possibly more than onecomplementary moiety binding site 331. The complementary moiety bindingsite 331 binds specifically to complementary moieties 335 exposed on thecomplementary surface 333. The complementary moiety binding peptide 327is bound to the complementary surface 333 because it is composed of atleast one complementary moiety binding domain 329 which contains atleast one complementary moiety binding site 331. In addition to thecomplementary moiety binding domain 329, the complementary moietybinding peptide 327 also contains at least one but possibly more thanone target domain 337. As discussed above, the target binding domain 311of the PP-binding peptide 305 binds to the target domain 337.

It should be clear to one skilled in the art that the complementarysurface 333 may be composed of PP itself and the complementary moietybinding domain 327 could be a PP binding domain. This embodiment isuseful because it provides an adhesive that is specific and functionaleven in adverse circumstances among such circumstances, as not limitingexamples, are the presence of water, oil, or dirt.

FIG. 5 depicts another embodiment of the invention useful for bindingtwo surfaces together. In this embodiment neither surface need tonecessarily contain PP, although that possibility is not excluded. Theprimary structure of the peptide based adhesive is similar to that showin FIG. 4. Two surfaces are provided 433, 439. Each surface comprising atarget molecule 435, 441 either of which may or may not be the same andmay or not be PP. A peptide diblock is provided comprising in each casea target binding peptide 405 with a target binding domain 411 comprisinga target binding site 431. The target binding peptide 405 comprises a PPbinding domain 407 having a PP binding site 415, useful for binding PPmoieties. Juxtapositioning of the two surfaces in the presence of PPmoieties 403 results in adhesion of the surfaces though the PP.

It will be apparent to the skilled person that this embodiment may alsobe practiced with the addition of a benefit agent(s) and/or peptidedomain(s) as describe above. This embodiment is useful because itprovides an adhesive that is specific and functional even in adversecircumstances among such circumstances, as not limiting examples, arethe presence of water, oil, or dirt.

FIG. 6 depicts an embodiment of the invention in which a surface 533 maybe coated with PP 503 using PP-binding peptide 505 containing a targetbinding domain 511. FIG. 6 panel A depicts a surface 533 coated with atarget peptide 527 that contains in part or whole a target domain 537.The target peptide 527 may be applied to the surface 533 by any methodeither described herein or known in the art; one method will bedescribed in detail later when discussing FIG. 6 panel D. The PP-bindingpeptides 505 used in this embodiment each contain, as described above,at least one PP binding domain 507 which in turn contains at least onePP binding site 515. The PP binding site 515 binds PP 503 specificallyas described elsewhere herein. In addition to the PP binding domain 507,the PP-binding peptides 505 also each contain at least one but possiblymore than one target binding domain 511. The target binding domain usedis selected, or created, using methods described or known, to bindspecifically to the target domain 537 of the target peptide 527 on thesurface 533. If the PP-binding peptide 505 and PP moieties 503 asdescribed are allowed to move freely in a medium around the exposedsurface 533, PP-binding peptide 505 will adhere to the peptides 527 onthe surface 533 through the bonding of the target binding domain 511 ofthe PP-binding peptide 505 to the target domain 537 of the surfacepeptide 527. PP moieties in the media will bind to the PP binding site515 of the PP binding domain 507 of the PP-binding peptide 505 forming adiblock structure. With PP 503 bound to the PP-binding peptide 505 andit in turn bound to the surface peptides 527 that are bound to thesurface 533, PP 503 moieties will coat the surface 533.

FIG. 6 panel B, depicts the same interactions of PP-binding peptide 505,PP 503 and a peptide coated surface 533, as described in panel A, withthe addition of a benefit agent 517 coupled to the PP-binding peptide505 which itself contains at least one target binding domain 511. Usingmethods described herein this embodiment couples a benefit agent 517 tothe PP-binding peptide 505. When the complex of the benefit agent 517and the PP-binding peptide 505 bind a PP moiety 503 a triblock isformed. The triblock structure does not prevent the benefit agent 517from being functionally active or from the target binding domain 511from binding the target peptide 527. The addition of a benefit agent 517to the PP-binding peptide 505 allows the surface to be coated with botha benefit agent 519 and PP moieties 503. Non-limiting examples ofbenefits agents 517 that may be used with this embodiment are dyes,colorants, antimicrobials, and stain repelling moieties.

FIG. 6 panel C, depicts the same interactions as in panel A, andprovides the addition of a benefit agent 523 bound to PP. In thisembodiment, the benefit agent 523 is attached to the PP moiety or bead503 with a bond structure 525. The bonding structure may be part of thebenefit agent 523 or may be an independent structure that is bound tothe PP 503 for the purpose of binding the benefit agent 523. Or in thealternative, the bond structure 525 may be bound to the benefit agent523 for the purpose of binding it to the PP 503. The binding structure523 may be a permanent bond, but in some forms of the embodiment may beeasily broken under certain conditions. In other forms of the embodimentthe binding structure 525 may allow the benefit agent 523 to be leachedfrom the PP 503 under certain conditions. In still other forms of theembodiment the binding structure might allow the benefit agent 523 to bereleased over time at regular or specific time intervals. In analternative form of this embodiment the binding structure 525 itself maybe in whole or in part be composed of PP. In this form, the PP 503 maybe in whole or in part the binding structure 525. The benefit agent 523may be partially or fully embedded with the PP 503. A triblock structureis formed when the benefit agent 523 coupled to the PP moiety 503 thatis inturn bound to the PP-binding peptide 505. The triblock structure iscapable of binding the target peptide as described above.

FIG. 6 panel D, Depicts a PP-binding peptide 505 and PP moiety or bead503 similar to the PP-binding peptide 505 described in panel A. In thisembodiment, the target peptide 527 depicted is not attached directly tothe surface 533. The target peptide 527 contains a target binding domain537 as in panels A, B, and C and additionally contains a surface moietybinding domain 529. The surface moiety binding domain 529 is selected tobind specifically to a known moiety that is known to be exposed on thesurface 533. The surface binding moiety domain 529 contains at leastone, but possibly more than one, surface moiety bind site 531. Thesurface moiety binding site 531 is the point of attachment between thesurface moiety 535 and the surface moiety binding domain 529. Throughthe interaction of the surface moiety binding domain 529 and the surfacemoiety 535 the PP-binding peptide 505 is attached to the surface 533.Further through the binding interaction of the PP moieties or beads 503and PP-binding peptide 505 bound to the surface 503, the surface 503 iscoated with PP moieties or beads 503.

EXAMPLES

The present invention is further defined in the following Examples. Itshould be understood that these Examples, while indicating preferredembodiments of the invention, are given by way of illustration only.From the above discussion and these Examples, one skilled in the art canascertain the essential characteristics of this invention, and withoutdeparting from the spirit and scope thereof, can make various changesand modifications of the invention to adapt it to various uses andconditions.

The meaning of abbreviations used is as follows: “min” means minute(s),“sec” means second(s), “h” means hour(s), “μL” means microliter(s), “mL”means milliliter(s), “L” means liter(s), “nm” means nanometer(s), “mm”means millimeter(s), “cm” means centimeter(s), “μm” means micrometer(s),“mM” means millimolar, “M” means molar, “mmol” means millimole(s),“μmole” means micromole(s), “g” means gram(s), “μg” means microgram(s),“mg” means milligram(s), “g” means the gravitation constant, “rpm” meansrevolutions per minute, “pfu” means plague forming unit, “BSA” meansbovine serum albumin, “ELISA” means enzyme linked immunosorbent assay,“IPTG” means isopropyl β-D-thiogalactopyranoside, “A” means absorbance,“A₄₅₀” means the absorbance measured at a wavelength of 450 nm, “TBS”means Tris-buffered saline, “TBST-X” means Tris-buffered salinecontaining Tween® 20 where “X” is the weight percent of Tween® 20,“Xgal” means 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside, “SEM”means standard error of the mean, “vol %” means volume percent.

General Methods:

Standard recombinant DNA and molecular cloning techniques used in theExamples are well known in the art and are described by Sambrook, J.,Fritsch, E. F. and Maniatis, T., Molecular Cloning: A Laboratory Manual,Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989, byT. J. Silhavy, M. L. Bennan, and L. W. Enquist, Experiments with GeneFusions, Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y., 1984,and by Ausubel, F. M. et al., Current Protocols in Molecular Biology,Greene Publishing Assoc. and Wiley-Interscience, N.Y., 1987.

Materials and methods suitable for the maintenance and growth ofbacterial cultures are also well known in the art. Techniques suitablefor use in the following Examples may be found in Manual of Methods forGeneral Bacteriology, Phillipp Gerhardt, R. G. E. Murray, Ralph N.Costilow, Eugene W. Nester, Willis A. Wood, Noel R. Krieg and G. BriggsPhillips, eds., American Society for Microbiology, Washington, D.C.,1994, or by Thomas D. Brock in Biotechnology: A Textbook of IndustrialMicrobiology, Second Edition, Sinauer Associates, Inc., Sunderland,Mass., 1989.

All reagents, restriction enzymes and materials used for the growth andmaintenance of bacterial cells were obtained from Aldrich Chemicals(Milwaukee, Wis.), BD Diagnostic Systems (Sparks, Md.), LifeTechnologies (Rockville, Md.), or Sigma Chemical Company (St. Louis,Mo.), unless otherwise specified.

Example 1 Selection of Polypropylene-Binding Peptides Using Biopanning

The purpose of this Example was to identify phage peptides that bind topolypropylene (PP) using a modified phage display biopanning method.

Biopanning—Phage Display Selection of PP-Binding Peptides

Phase Display Peptide Libraries:

The phage libraries used in the present invention, Ph.D.-12™ PhageDisplay Peptide Library Kit and Ph.D.-7™ Phage Display Library Kit, werepurchased from New England BioLabs (Beverly, Mass.). These kits arebased on a combinatorial library of random peptide 7 or 12-mers fused toa minor coat protein (pill) of M13 phage. The displayed peptide isexpressed at the N-terminus of pill, such that after the signal peptideis cleaved, the first residue of the coat protein is the first residueof the displayed peptide. The Ph.D.-7 and Ph.D.-12 libraries consist ofapproximately 2.8×10⁹ and 2.7×10⁹ sequences, respectively. A volume of10 μL contains about 55 copies of each peptide sequence. Each initialround of experiments was carried out using the original library providedby the manufacturer in order to avoid introducing any bias into theresults.

Biopanning Against a PP Surface:

The polypropylene-binding peptides were identified using the biopanningmethod described below. The polypropylene substrate used in thebiopanning method was a polypropylene mesh material, specifically,Hernia Repair/Reconstructive Surgery Prosthetics Patch (obtained fromBARD Medical, Davol Inc., Cranston, R.I.). The polypropylene mesh wascut into 1-cm squares and pretreated with 90% isopropanol for 30 min atroom temperature, followed by washing 5 times for 10 min each withdeionized water before the panning process.

The mesh was placed in a tube and 5 mL of blocking buffer consisting of1 mg/mL BSA in TBST containing 0.5% Tween® 20 (TBST-0.5%) was added tothe tube and incubated for 1 h at 4° C. The PP mesh was washed 5 timeswith TBST-0.5% and then 2 mL of TBST-0.5% containing 1 mg/mL BSA wasadded to each tube. Then, 10 μL of the original phage library (2×10¹¹pfu), either the 12-mer or 7-mer library, was added to the PP mesh andincubated for 15 min at room temperature. The PP mesh was washed 10times with TBST-0.5%. The PP mesh was then transferred to a clean tube,2 mL of a non-specific elution buffer consisting of 1 mg/mL BSA in 0.2 Mglycine-HCl, pH 2.2, was added to the tube and incubated for 10 min. ThePP mesh was washed three more times with the elution buffer and thenwashed three times with TBST-0.5%. The PP mesh, which had acid resistantphage peptides still attached, was used to directly infect the hostcells E. coli ER 2738 (New England BioLabs, Beverly, Mass.), for phageamplifications. The PP mesh was incubated with an overnight E. coliER2738 culture diluted 1:100 in LB medium, at 37° C. for 4.5 h. Afterthis time, the cell culture was centrifuged for 30 sec and the upper 80%of the supernatant was transferred to a fresh tube, ⅙ volume of PEG/NaCl(20% polyethylene glycol-800, obtained from Sigma Chemical Co. St.Louis, Mo., 2.5 M sodium chloride) was added, and the phage was allowedto precipitate overnight at 4° C. The precipitate was collected bycentrifugation at 10,000×g at 4° C. and the resulting pellet wasresuspended in 1 mL of TBS. This was the first round of amplified stock.The amplified first round phage stock was then titered according to themethod described below. For the next round of biopanning, more than2×10¹¹ pfu of phage stock from the first round was used. The biopanningprocess was repeated for 4 rounds.

After the acid wash steps in the final round of biopanning, the PP meshwas used to directly infect 500 μL of mid-log phase bacterial hostcells, E. coli ER2738, which were then grown in LB medium for 20 min andthen mixed with 3 mL of agarose top (LB medium with 5 mM MgCl₂, and 0.7%agarose) at 45° C. This mixture was spread onto a LB medium/IPTG/S-Gal™plate (LB medium with 15 g/L agar, 0.05 g/L IPTG, and 0.04 g/L S-Gal™)and incubated overnight at 37° C. The black plaques were counted tocalculate the phage titer. The single black plaques were randomly pickedfor DNA isolation and sequencing analysis.

A total of four rounds of biopanning were performed and the amino acidsequences of the high affinity, polypropylene-binding phage peptides aregiven in Table 7.

TABLE 7 Amino Acid Sequences of High AffinityPP-Binding Phage Peptides from the 7- and 12-Mer LibrariesAmino Acid Sequence SEQ ID NO: TSDIKSRSPHHR 1 HTQNMRMYEPWF 2 LPPGSLA 3MPAVMSSAQVPR 4 NQSFLPLDFPFR 5 SILSTMSPHGAT 6 SMKYSHSTAPAL 7

1. A peptide reagent having a general structure selected from the group consisting of: a) PP_(m)-(PPBP-BAp)n; and b) PP_(m)-(PPBP-L-BA)n; wherein: i) PP is a polypropylene moiety ii) PPBP is a polypropylene binding peptide having a polypropylene binding domain; iii) BA is at least one benefit agent wherein the at least one benefit agent is a colorant; iv) L is a linker molecule; v) m=the number of polypropylene moieties available for binding, and m is at least one; vi) n=m; and vii) p=1-20.
 2. A peptide reagent according to claim 1 wherein the polypropylene binding peptide comprises a polypropylene binding sequence selected from the group consisting of SEQ ID NO's: 1-7.
 3. A peptide reagent according to claim 1 wherein the linker molecule is selected from the group consisting of: a peptide linker, and an organic linker.
 4. A peptide reagent according to claim 1 wherein the polypropylene moiety is incorporated into a surface.
 5. A peptide reagent according to claim 4 wherein the surface is selected from the group consisting of a solid support, a bead, a microsphere, a sheet, and a fiber.
 6. A peptide reagent according to claim 5 wherein the microsphere comprises a dye.
 7. A peptide reagent according to claim 4 wherein the surface is coated on a secondary surface.
 8. A peptide reagent according to claim 1 wherein the polypropylene moiety is comprised within a polypropylene film.
 9. A peptide reagent according to claim 1 wherein the colorant is a hair colorant selected from the group consisting of D&C Yellow 1 and 3, HC Yellow 6 and 8, D&C Blue 1, HC Blue 1, HC Brown 2, HC Red 5, 2-nitro-paraphenylenediamine, N,N-hydroxyethyl-2-nitro-phenylenediamine, 4-nitro-indole, iron oxides, and carbon black.
 10. A peptide reagent according to claim 1 wherein the polypropylene binding peptide is isolated by a process comprising the steps of: (i) providing a library of combinatorially generated peptides; (ii) contacting the library of (i) with a polypropylene sample to form a reaction solution comprising: (A) peptide-polypropylene complex; (B) unbound polypropylene, and (C) uncomplexed peptides; (iii) isolating the peptide-polypropylene complex of (ii); (iv) eluting the weakly bound peptides from the isolated peptide complex of (iii) whereby the polypropylene binding peptide is isolated. 